Expression and interaction analysis of Arabidopsis Skp1-related genes

被引:69
作者
Takahashi, N
Kuroda, H
Kuromori, T
Hirayama, T
Seki, M
Shinozaki, K
Shimada, H
Matsui, M [1 ]
机构
[1] RIKEN, Yokohama Inst, Genom Sci Ctr,Tsurumi Ku, Plant Funct Explorat Team,Plant Funct Genom Res G, Yokohama, Kanagawa 2300045, Japan
[2] Tokyo Univ Sci, Dept Biol Sci & Technol, Noda, Chiba 2788510, Japan
[3] RIKEN, Yokohama Inst, Plant Mutat Explorat Team, Plant Funct Genom Res Grp,Tsurumi Ku, Yokohama, Kanagawa 2300045, Japan
[4] RIKEN, Tsukuba Inst, Plant Mol Biol Lab, Tsukuba, Ibaraki 3050074, Japan
[5] Yokohama City Univ, Grad Sch Integrated Sci, Tsurumi Ku, Yokohama, Kanagawa 2300045, Japan
关键词
Arabidopsis thaliana; ASK; F-box protein; GUS; protein degradation; yeast two-hybrid system;
D O I
10.1093/pcp/pch009
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Specific protein degradation has been observed in several aspects of development and differentiation in many organisms. One example of such proteolysis is regulated by protein polyubiquitination that is promoted by the SCF complex consisting of Skp1, cullin, and an F-box protein. We examined the activities of the Arabidopsis Skp1-related proteins (ASKs). Among 19 annotated ASK genes, we isolated 16 of the corresponding cDNAs (ASK1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 16, 17, 18, 19), and examined their gene products for interactions with 24 representatives of F-box proteins carrying various classes of the C-terminal domains using the yeast two-hybrid system. As a result, we found diverse binding specificities: ASK1, ASK2, ASK11 and ASK12 interacted well with COI1, FKF1, UFO-like protein, LRR-containing F-box proteins, and other F-box proteins with unknown C-terminal motifs. We also observed specific interaction between F-box proteins and ASK3, ASK9, ASK13, ASK14, ASK16 and ASK18. In contrast, we detected no interaction between any of the 12 ASK proteins and F-box proteins containing CRFA, CRFB or CRFC domains. Both histochemical and RT-PCR analysis of eight ASK genes expression revealed unique expression patterns for the respective genes.
引用
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页码:83 / 91
页数:9
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